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Artykuły w czasopismach na temat "MAB PURIFICATION"
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Shinagawa, Kunihiro, Katsuhiko Omoe, Naonori Matsusaka i Shunji Sugii. "Immunological studies on staphylococcal enterotoxin D: production of murine monoclonal antibodies and immunopurification". Canadian Journal of Microbiology 37, nr 8 (1.08.1991): 586–89. http://dx.doi.org/10.1139/m91-099.
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Eight murine monoclonal antibodies (MAbs) against staphylococcal enterotoxin D (SED) were obtained by fusion of myeloma cells with mouse spleen cells immunized with SED only or a combination of SED and either enterotoxin A (SEA) or enterotoxin E (SEE). When only SED was used as an immunogen, six MAbs were specific for SED only, whereas one MAb was reactive with both SED and SEE when both SEs were used as immunogens. One MAb reacted with SEA, SED, and SEE when both SEA and SED were used as immunogens. A MAb with the highest reactivity to SED was used to prepare an immunosorbent for purification of SED by immunoaffinity chromatography. Approximately 70% of the partially purified SED was recovered in the eluate. The purified SED was electrophoretically and antigenically pure. Immunoaffinity chromatography proved useful in the purification of SED in terms of ease of purification, percent enterotoxin, and enterotoxin purity. Key words: enterotoxin D, monoclonal antibodies, Staphylococcus aureus.
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Shi, Wenshu, Haiyang Hao, Mengran Li, Jianqin Niu, Yaning Hu, Xingbo Zhao i Qiuyan Li. "Expression and Purification of a PEDV-Neutralizing Antibody and Its Functional Verification". Viruses 13, nr 3 (12.03.2021): 472. http://dx.doi.org/10.3390/v13030472.
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Porcine epidemic diarrhea virus (PEDV) is a highly infectious and pathogenic virus causing high morbidity and mortality, especially in newborn piglets. There remain problems with contemporary PEDV vaccines, in part because of the rapid variation of PEDV, poor conferred immunity, and numerous side effects. The ability to produce PEDV-neutralizing antibodies suggests that we may be able to increase the success rate of PEDV prevention in piglets using these antibodies. In this study, we produced an anti-PEDV S protein monoclonal antibody (anti-PEDV mAb-2) that neutralized PEDV-CV777 (a G1 strain), PEDV-SDSX16 and PEDV-Aj1102 (two G2 strains). In vivo challenge experiments demonstrated that anti-PEDV mAb-2 inhibited the PEDV infection in piglets. We also produced three HEK293 cell lines that expressed anti-PEDV mAb-2. Overall, our study showed that anti-PEDV mAb-2 produced from hybridoma supernatants effectively inhibited PEDV infection in piglets, and the recombinant HEK293 cell lines expressed anti-PEDV mAb-2 genes.
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Deplazes, P., i B. Gottstein. "A monoclonal antibody against Echinococcus multilocularis Em2 antigen". Parasitology 103, nr 1 (sierpień 1991): 41–49. http://dx.doi.org/10.1017/s0031182000059278.
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A monoclonal antibody (MAb G11) species-specific to the Em2 antigen of Echinococcus multilocularis was generated for (i) further biological characterization of the Em2 antigen, (ii) easy affinity-purification of Em2 antigen for immunodiagnostic and immunological investigations and (iii) development of a sandwich-ELISA for the detection of Em2 antigen in diagnostic samples and thus species-specific identification of E. multilocularis metacestode material. The MAb G11 was used in an antibody sandwich-ELISA to detect soluble Em2 antigen with a methodical sensitivity of 80 ng E. multilocularis antigen/ml of solution. MAb G11 specifically detected Em2 antigen in all of 15 E. multilocularis-isolates originating from various geographical areas and in none of other helminth isolates (e.g. Echinococcus granulosus, E. vogeli, and others). Further biological analysis by FITC-labelled MAb G11 demonstrated unique binding activity to the laminated layer of the metacestode. Also, oncospheres were binding FITC-labelled MAb G11 on an outer layer synthesized during cultivation in vitro for 13 days after hatching. Application of the MAb G11 antibody sandwich-ELISA for investigation of solubilized oncospheres confirmed the in vitro synthesis of Em2 antigen by oncospheres on day 13 p.i. Adult stages (somatic antigens) and freshly hatched oncospheres were always MAb G11 negative. Solid-phase MAb G11 was used for purification of the corresponding Em2 antigen by affinity chromatography. A preliminary serological evaluation of the Em2(G11) antigen by ELISA revealed identical immunodiagnostic characteristics, compared to Em2 obtained by classical means, thus suggesting the presented method for future isolation of large-scale Em2 antigen.
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Dasch, J. R., D. R. Pace, W. Waegell, D. Inenaga i L. Ellingsworth. "Monoclonal antibodies recognizing transforming growth factor-beta. Bioactivity neutralization and transforming growth factor beta 2 affinity purification." Journal of Immunology 142, nr 5 (1.03.1989): 1536–41. http://dx.doi.org/10.4049/jimmunol.142.5.1536.
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Abstract Four mAb able to recognize transforming growth factor-beta 2 (TGF-beta)2 were obtained. One of these mAb, 1D11.16, was able to neutralize the biological activity of both TGF-beta 1 and beta 2 in vitro. This was demonstrated in an Il-1, PHA-dependent thymocyte mitogenic assay that is inhibitable by TGF-beta in a dose-dependent manner. All four mAb recognized the dimeric form of TGF-beta 2 in Western blots. The mAb were also found to immunoprecipitate [125I]-TGF-beta 2. mAb 3C7.14 coupled to Sepharose could efficiently immunoaffinity purify TGF-beta 2 from a complex mixture of proteins. Affinity constants were determined for the four mAb and they ranged from 3.4 x 10(8) to 1.6 x 10(7) L/mol.
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Ziltener, H. J., I. Clark-Lewis, B. Fazekas de St Groth, P. C. Orban, L. E. Hood, S. B. Kent i J. W. Schrader. "Monoclonal antipeptide antibodies recognize IL-3 and neutralize its bioactivity in vivo." Journal of Immunology 140, nr 4 (15.02.1988): 1182–87. http://dx.doi.org/10.4049/jimmunol.140.4.1182.
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Abstract Mixtures of synthetic peptides corresponding to segments of murine IL-3 or synthetic IL-3 were used to raise murine mAb. Several mAb able to recognize synthetic IL-3 were obtained, two of which exhibited significant cross-reactivity with native IL-3 as shown by precipitation of biosynthetically 35S-labeled IL-3 and their effectiveness as affinity reagents for the purification of IL-3 from conditioned medium. The amino acid sequence recognized by the two mAb was determined by using synthetic peptide segments of IL-3. In both cases binding of the mAb to synthetic IL-3 was inhibited best with a hexapeptide corresponding to the amino acid residues 130-135 of IL-3, although the mAb differed in other characteristics. Neither mAb neutralized IL-3 bioactivity in vitro. However, we observed that in vivo administration of one mAb abrogated the increase in splenic mast cells and their precursors that normally occurred in mice bearing a s.c. IL-3-producing tumor, WEHI-3B.
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Harada, R., N. Okada, T. Fujita i H. Okada. "Purification of 1F5 antigen that prevents complement attack on homologous cell membranes." Journal of Immunology 144, nr 5 (1.03.1990): 1823–28. http://dx.doi.org/10.4049/jimmunol.144.5.1823.
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Abstract A mAb, 1F5, has the ability to cause hemolysis by human serum of human E treated with neuraminidase via the alternative C pathway. By Western blotting, this mAb reacts with a glycoprotein having a molecular mass of 20 kDa (1F5Ag). 1F5Ag was isolated from human E by affinity chromatography with mAb-coupled Sepharose. Purified 1F5Ag was then adsorbed to guinea pig E rendering them resistant to human C attack by both the classical and the alternative pathways. Furthermore, experiments with isolated C components revealed that 1F5Ag interferes with both homologous human C8 and C9 in the terminal stage of the C reaction, whereas it has little effect on hemolysis by rabbit C8 and C9. Therefore, 1F5Ag can be called HRF20, which stands for homologous restriction factor of 20 kDa.
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Kruse, Thomas, Axel Schmidt, Markus Kampmann i Jochen Strube. "Integrated Clarification and Purification of Monoclonal Antibodies by Membrane Based Separation of Aqueous Two-Phase Systems". Antibodies 8, nr 3 (2.07.2019): 40. http://dx.doi.org/10.3390/antib8030040.
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Therapeutic monoclonal antibodies (mAb) are used for the treatment of numerous serious diseases, which have led to an increasing demand over the last decades. Increased cell density and mAb titer of the cultivation broth lead to great challenges for the subsequent clarification and capture operations in the downstream process. As an alternative approach to the conventional downstream process, a selective mAb extraction via an aqueous two-phase system (ATPS) directly from the cultivation broth of a mAb producing industrial relevant chinese hamster ovary (CHO) cell line was investigated. An efficient purification of the mAb was accomplished by the ATPS composition. The phase separation was realized by a newly developed membrane based phase separator. Moreover, a complete cell removal was integrated into this process by the used membrane. A selectivity between both phases was achieved by membrane modification. Yields up to 93% in the light phase and removal of process related impurities were obtained after aqueous two-phase extraction (ATPE). Phase separation performance as well as contact angles on the membrane were characterized for different ATPS. ATPE directly from the cultivation broth in combination with the new membrane based phase separation led to a mAb yield of 78% with a simultaneous reduction of deoxyribonucleic acid (DNA) and host cell protein (HCP) load.
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Wei, Yuping, Jiandong Xu, Liang Zhang, Yankai Fu i Xia Xu. "Development of novel small peptide ligands for antibody purification". RSC Advances 5, nr 82 (2015): 67093–101. http://dx.doi.org/10.1039/c5ra07829f.
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Small peptide ligands which were designed based on the interactions with human immunoglobulin G (IgG) using the molecular simulations, can offer a potential alternative for mAb purification with elution condition at pH 9 and pH 3.
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Santos, Carlos F., Andrew S. Greene, Maria Cristina O. Salgado i Eduardo B. Oliveira. "Conversion of renin substrate tetradecapeptide to angiotensin II by rat MAB elastase-2". Canadian Journal of Physiology and Pharmacology 82, nr 11 (1.11.2004): 1000–1005. http://dx.doi.org/10.1139/y04-102.
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A new approach for the purification of rat mesenteric arterial bed (MAB) elastase-2 has been developed using the chromogenic substrates N-succinyl-Ala-Ala-Pro-Phe-p-nitroanilide and N-succinyl-Ala-Ala-Pro-Leu-p-nitroanilide to monitor the enzymatic activity during various stages of purification. The purified enzyme was evaluated in the presence of various inhibitors and confirmed to have angiotensin (Ang) II-forming ability. The active site-directed inhibitor acetyl-Ala-Ala-Pro-Leu-chloromethylketone (100 µmol·L-1), described for human pancreatic elastase-2, abolished the enzymatic activity, confirming that the enzyme is an elastase-2. Chymostatin (100 µmol·L-1), an inhibitor regarded as selective for chymases, also showed a remarkable inhibitory effect (94%), whereas captopril (100 µmol·L-1) had no effect at all on the Ang II-forming activity. The Ang II precursor renin substrate tetradecapeptide (RS-14P) was converted into Ang II by the rat MAB elastase-2 with the following kinetic constants: Km = 124 ± 21 µmol·L-1; Kcat = 629 min-1; catalytic efficiency (Kcat /Km) = 5.1 min-1 µ(mol/L)-1. In conclusion, the strategy for the purification of rat MAB elastase-2 with the chromogenic substrates proved to be simple, rapid, accurate, and highly reproducible; therefore, it can be reliably and conveniently used to routinely purify this enzyme. The kinetic parameters for the formation of Ang II from RS-14P by rat MAB elastase-2 emphasize differences in substrate specificity between this and other Ang II-forming enzymes.Key words: N-succinyl-Ala-Ala-Pro-Phe-p-nitroanilide, N-succinyl-Ala-Ala-Pro-Leu-p-nitroanilide, elastase-2, angiotensin II, renin substrate tetradecapeptide.
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Valdés, Rodolfo, Tatiana Álvarez, Andrés Tamayo, Eutimio G. Fernandez, José Montero, Déborah Geada, William Ferro i in. "New Mab CB.Hep-1 Purification Process Eliminates the Need for Pre-Chromatographic Purification. Stability Demonstrated Over 100 Purification Cycles". Chromatographia 67, nr 11-12 (18.04.2008): 923–27. http://dx.doi.org/10.1365/s10337-008-0607-5.
Pełny tekst źródłaRozprawy doktorskie na temat "MAB PURIFICATION"
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Fernando, Samantha. "Monoclonal antibody (mAb) purification by counter current chromatography (CCC)". Thesis, Brunel University, 2011. http://bura.brunel.ac.uk/handle/2438/6522.
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Counter current chromatography (CCC) is a form of liquid liquid chromatography, which the Brunel Institute for Bioengineering (BIB) team have developed to process scale. In this thesis, its application has been successfully extended to the rapid, scalable purification of monoclonal antibodies (mAb) from mammalian cell culture, using aqueous two-phase systems (ATPS) of inorganic salts and polymer. A polyethylene glycol (PEG) and sodium citrate system was found to be the most appropriate by robotic phase system selection. The search for an economical alternative to protein A HPLC is a substantial bioprocessing concern; in this work CCC has been investigated. Initial studies showed that unpredictably, despite separation from impurities being achieved, some loss in the IgG‘s ability to bind to Protein A was seen, as confirmed by Protein A BiaCore analysis. CCC machines were seen to adversely affect IgG functionality. This led to a systematic investigation of the effect of CCC phase mixing on IgG functionality in a number of different CCC instruments, allowing direct comparisons of modes of CCC (hydrodynamic and hydrostatic CCC) and their associated mixing (wave-like and cascade, respectively). The varying g forces produced within the CCC column were determined using a recently developed model to calculate g force range. The effect of interfacial tension was also studied using a custom built 'g' shaker. The optimum CCC mode was identified to be the non synchronous CCC, operated in a hydrodynamic mode but allowing bobbin to rotor speed (Pr ratio) to be controlled independently. In a normal synchronous J type centrifuge a Pr of 1 is fixed, this is where the bobbin and rotor speed are identical I.e. one bobbin rotation (where mixing occurs) to one rotor revolution (where settling occurs). Constraints were seen with this 1:1 ratio and the separation of mAb using ATPS. This work has shown with the use of the non synchronous CCC at a Pr of 0.33, mixing is reduced and rotor rotations increased. Consequently the associated g force range is decreased. Furthermore, by the extension of settling time, the clear separation of the mAb from impurities has been achieved with retention of biological activity. This thesis demonstrates the importance of settling time for ATPS in phase separation and documents the fundamental requirements for the successful separation of biologics. Purified non synchronous CCC samples have additionally undergone rigorous quality control testing at Lonza Biologics by their purification scientists. This work has ultimately showed that with optimisation, the non synchronous CCC can be used to produce biological samples that are of industry standard.
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Holzberg, David Alexander. "Untersuchungen zur spezifischen Funktion von JNK-Signalwegen mit Hilfe von zellpermeablen Peptiden und tandem affinity purification". [S.l. : s.n.], 2004. http://deposit.ddb.de/cgi-bin/dokserv?idn=972264175.
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A, S. Jijumon. "Systematic characterization of a large number of Microtubule-Associated Proteins using purification-free TIRF-reconstitution assays Purification of tubulin with controlled post-translational modifications by polymerization–depolymerization cycles Microtubule-Associated Proteins: Structuring the Cytoskeleton Purification of custom modified tubulin from cell lines and mouse brains by polymerization-depolymerization cycles". Thesis, université Paris-Saclay, 2021. http://www.theses.fr/2021UPASL007.
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Le cytosquelette des microtubules (MTs) est constitué de filaments dynamiques impliqués dans une multitude de fonctions telles que la division cellulaire, le maintien de forme des cellules, les battements ciliaires ou encore la différenciation neuronale. Une régulation stricte des fonctions des MTs est donc d'une grande importance pour l'homéostasie cellulaire, et toute perturbation pourrait potentiellement conduire à des maladies comme le cancer, les ciliopathies ou la neurodégénérescence. Dans un contexte cellulaire, les propriétés des MTs peuvent être contrôlées par leurs interactions avec une grande variété de protéines associées (MT-associated proteins ; MAPs). Notre connaissance de ces interacteurs s'est continuellement enrichie au cours des dernières décennies, mais il n'existe à ce jour aucune étude systématique visant à décrire et à classer ces protéines en fonction de leurs mécanismes de liaison et de leurs effets structuraux sur les MTs. Dans mon projet de thèse, j’ai mis au point un essai permettant une analyse rapide et systématique à la base des lysats clarifiés de cellules humaines surexprimant une multitude des différents MAPs. Le comportement dynamique des MT en présence d'environ 50 MAPs différentes a été imagé à l'aide de la microscopie TIRF. Cela nous permet d'étudier le comportement des MAP dans une situation proche de leur environnement naturel, mais en éliminant la complexité de l'espace intracellulaire, telle que l'encombrement par des organelles et des filaments du cytosquelette à l'intérieur de l'espace intracellulaire confiné. En effet, la plupart des MAPs étaient bien solubles dans notre approche d'extraction, tandis que les approches de purification pour plusieurs d'entre elles ont conduit à leur précipitation, rendent les expériences de reconstitution in vitro classique impossible. Ma nouvelle approche m’a permis de définir plusieurs nouvelles protéines comme de véritables MAP. J’ai montré que des MAPs non-caractérisées auparavant ont des effets étonnamment différents sur la polymérisation et la structure des MTs, créant ainsi une variété de réseaux de MT distincts. J’ai également démontré que mon approche permet d'étudier les structures des MAPs associées aux MTs par cryo-microscopie électronique, ou d'étudier le dynamique des MTs porteuses de mutations trouvées dans les pathologies humaines. J’ai également démontré que mon approche permet à tester la sensibilité des MAPs aux modifications post-traductionnelles de la tubuline, ou d'étudier le rôle des MAPs dans les interactions entre l'actine et les MTs. Mon approche expérimentale permet donc de mieux comprendre comment les MAP et les MT contrôlent ensemble le fonctionnement du cytosqueletteMicrotubules (MTs) are dynamic filaments involved in a plethora of functions such as cell division, cell shape, ciliary beating, neuronal differentiation. Strict regulation of MT functions is therefore of high importance for the cellular homeostasis, and any perturbations could potentially lead to diseases like cancer, ciliopathies and neurodegeneration. At the protein level, there are accumulating studies showing that MT properties can be controlled via interaction with a large variety of MT-associated proteins (MAPs). Our knowledge of MAPs has been enriched over time, but up to this date no systematic studies exist that aim to describe and categorize these proteins according to their binding mechanisms and structural effects on MTs. In my PhD project, I have developed an assay for rapid and systematic analysis of MAPs using cleared lysates of cultured human cells in which I overexpress a variety of different MAPs. The dynamic behaviour of growing MTs in the presence of those MAPs were imaged using TIRF microscopy. This allows me to study the behaviour of around 50 MAP candidates in a situation close to their natural environment, but eliminating complexity coming from different organelles and crammed cytoskeleton filaments inside the confined intracellular space. Indeed, most MAPs were nicely soluble in the extract approach, while purification attempts of several of them led to protein precipitation, thus making classical invitro reconstitution approaches impossible. This novel approach allowed me to compare many MAPs under similar experimental conditions, and helped to define several novel proteins as bona-fide MAPs. I demonstrate that previously uncharacterized MAPs have strikingly different effects on MT polymerization and MT structure, thus creating a variety of distinct MT arrays. I further extended this cell-free pipeline to study structures of MAPs bound to MTs by cryo-electron microscopy, or to study the MT interactions of MAPs carrying patient mutations. Finally, I demonstrated that my approach can be used to test the sensitivity of MAPs to tubulin PTMs, as well as to study the role of MAPs in actin-MT crosstalk. In the future, this novel approach will allow for a better mechanistic understanding of how MAPs and MTs together control cytoskeleton functions
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Moreno, Iglesias Ana. "Caracterización de la MAP kinasa ERK5 en células neuronales: papel en la isquemia cerebral e identificación de proteínas asociadas mediante tandem affinity purification". Doctoral thesis, Universitat Autònoma de Barcelona, 2009. http://hdl.handle.net/10803/3599.
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La vía de señalización celular MEK5-ERK5 juega un papel importante en sistema nervioso, dado que se activa frente a neurotrofinas, frente al estrés oxidativo producido por reactive oxygen species (ROS), tras la isquemia cerebral y tiene un papel en la especificación de fenotipo neuronal. Sin embargo, poco se conoce sobre los sustratos y proteínas que interaccionan con ERK5. En este trabajo se han identificado proteínas que interaccionan con ERK5 en células de neuroblastoma SH-SY5Y mediante el método TAP (Tandem-Affinity Purification). Se han obtenido diversas proteínas que incluyen proteínas de citoesqueleto, chaperonas y proteínas de metabolismo celular. Entre las proteínas identificadas destacan la proteína chaperona Hsp90β y la piruvato kinasa PKM2. Por otro lado, se ha determinado el papel que juega ERK5 en la isquemia cerebral, utilizando el modelo de privación de oxígeno y glucosa (OGD) en cultivos mixtos de neuronas corticales. ERK5 se degrada rápidamente en respuesta a la OGD en cultivos mixtos de neuronas corticales. Esta degradación es mediada por calpaína. ERK5 también se degrada rápidamente in vivo (4 horas) en córtex de cerebros de ratas Sprague-Dawley sometidas a isquemia por oclusión de la arteria cerebral media. En este trabajo se establece que tras la isquemia, la entrada masiva de calcio a través de los receptores de NMDA y la consiguiente activación de calpaína conlleva la degradación de ERK5, produciéndose así una disminución de los niveles totales de esta kinasa.
MEK5-ERK5 pathway plays an important role in nervous system. This pathway is activated in response to neutrophins, oxidative stress induced by reactive oxygen species (ROS), after cerebral ischemia and plays an important role in neuronal fate determination. However, little is known about substrates and proteins that interact with ERK5. This work describes the search for proteins which interact with ERK5 in the cell line SH-SY5Y using Tandem-Affinity Purification (TAP). Using this technique, we obtained different proteins that include citosqueleton components, chaperons and cell metabolism proteins, among which it is worth highlighting Hsp90β and piruvate kinase PKM2.
On the other hand, we have established the role of ERK5 in cerebral ischemia, using oxygen and glucose deprivation model (OGD) in rat mixed neuronal cortical cultures. Using this model we observed that ERK5 is quickly degraded in response to OGD, and that calpain is responsible for ERK5 degradation. ERK5 is also quickly (4 hours) degraded in vivo in cerebral cortex from Sprague-Dawley rats subjected to middle cerebral artery occlusion. On this work we establish that after cerebral ischemia, massive income of calcium mediated by NMDA receptors and resulting activation of calpain entails ERK5 degradation, producing a decrease in total levels of this kinase.
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Garcia, Clara Zeferino [UNIFESP]. "“De mal com o espelho”: um estudo sobre a (re)configuração de corpos femininos pela cirurgia plástica". Universidade Federal de São Paulo (UNIFESP), 2013. http://repositorio.unifesp.br/11600/41746.
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O trabalho investiga como se dá a (re)configuração corporal pela cirurgia plástica estética, a partir da perspectiva de um grupo específico de mulheres, que realizaram plásticas de lipoaspiração e mamoplastia de aumento. Trata-se de um grupo relativamente homogêneo, no que diz respeito à faixa etária e classe social, e que compartilha um mesmo ideal de mulher bela, bem sucedida e independente. A partir da afirmação da identidade de poderosa, uma concepção específica de feminilidade é valorizada pelo grupo, diretamente vinculada a um suposto poder de sedução feminino. Dentro do grupo, compartilham-se também valores advindos da moral da saúde, que a partir da designação de “bons” e “maus” hábitos, fornece critérios de avaliação individual à sociedade moderna contemporânea. Assim, a hexis corporal da poderosa é portadora de marcas distintivas que a remetem diretamente ao estilo de vida saudável, tido como moralmente superior. Nesse contexto, onde a gordura corporal localizada é vista como impura, a plástica de lipoaspiração é (re)significada como parte de um rito de purificação. Ao se retirar a gordura acumulada no corpo, busca-se apagar a memória de um passado de transgressões à moral da saúde e, mais especificamente, às normas instituídas sobre a alimentação e o trabalho. Por sua vez, a mamoplastia de aumento também não se mostra como um fato isolado nas trajetórias das pesquisadas, mas como uma prática vinculada a uma ascensão na estrutura social, vivida a partir da conquista de independência financeira, e mudança para territórios urbanos mais desenvolvidos. O poder aquisitivo, somado ao cumprimento de requisitos implícitos nas normas de purificação corporal do grupo, direciona essas mulheres à prática de “colocar silicone”, que pode ser entendida como parte de um rito de instituição, na medida em que vem para consagrar a inclusão no grupo valorizado das poderosas, marcando as diferenças entre essa categoria de mulher, tipicamente moderna, e as que ficam de fora: a “gorda”, a “feia” e a “pobre”.
The research investigates how the body (re)configuration by cosmetic plastic surgeries happens, according to the perspective of a specific group of women, who have undergone through plastic surgeries as liposuction and breast augmentation. It is a relatively homogeneous group, regarding to age and social class, and that shares the same ideal of beautiful, successful and independent woman. By the identity affirmation of poderosa (powerful woman), a specific conception of femininity is prized by the group, which is directly linked to the presumed female seductiveness. Within the group, are also shared values related to the health’s moral, or healthism, which defines "good" and "bad" habits, providing standards for individual evaluation in the contemporary modern society. Thus, the poderosa’s body hexis carries distinctive marks that relate it directly to a healthy lifestyle, seen as morally superior. In this context, where the concentrated body fat is seen as impure, the liposuction is (re)interpreted as part of a purification rite. By eliminating the accumulated fat in the body, it’s intended to erase the memory of a past of transgressions against the health’s moral, or healthism, and more specifically, against the rules that control food and work. At its side, the breast augmentation shows not to be an isolated incident in the trajectories studied, but a practice linked to a rise in the social structure, described like financial independence achievement, and moving to more developed urban areas. The purchasing power, combined with the fulfillment of requirements implied in the group standards of body purification, leads these women to the practice of "putting silicone" which can be understood as part of a rite of institution, once it comes to consecrate the inclusion into the valued group of poderosas, marking the differences between this category of woman, typically modern, and those left out: the "fat", the "ugly" and the "poor".
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Ducasse-Cabanot, Stéphanie. "MabA, β-cétoacyl-ACP réductase de mycobactérium tuberculosis : propriétés fonctionnelles et structurales et inhibition par l'antibiotique antituberculeux isoniazide". Toulouse 3, 2002. http://www.theses.fr/2002TOU3A201.
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Romir, Johannes [Verfasser]. "Expression and Purification of Arabidopsis Two-Component System Proteins, Diffraction Experiments with Crystals of Amide Synthetase NovL and Structure Determination of Human MAP Kinase p38 in Complex with Inhibitors / Johannes Romir". München : Verlag Dr. Hut, 2012. http://d-nb.info/1022535234/34.
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Huet, Moulard Émilie. "Étude de la régulation des récepteurs de peptides N-formyles". Université Joseph Fourier (Grenoble), 2007. http://www.theses.fr/2007GRE10077.
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Les cellules phagocytaires constituent la première ligne de défense contre les pathogènes. Leur migration dirigée vers le site infectieux et leurs fonctions microbicides sont l'aboutissement de voies de signalisation intracellulaires sollicitées par la stimulation de récepteurs couplés aux protéines G, les récepteurs de chimioattractants. Après fixation du ligand et transmission du signal par la protéine G, les récepteurs sont phosphorylés et interagissent avec les b-arrestines, protéines d'échafaudage concourrant à l'internalisation des récepteurs. Plusieurs exemples récents suggèrent que les b-arrestines pourraient également participer à la signalisation. Le travail présenté dans ce mémoire concerne les récepteurs de la famille FPR (Formyl Peptide Receptor) et plus spécialement le récepteur FPRL1 (FPR-like 1), pour lesquels de nouveaux agonistes dérivant de protéines bactériennes ou mitochondriales humaines ont été identifiés. La phosphorylation du récepteur FPRL1 a été caractérisée. Il a été montré que les -arrestines interagissent avec celui-ci et qu'elles sont indispensables à son internalisation. Diverses approches ont conclu que l'activation rapide des MAP kinases ERK1/2, enclenchée par la stimulation du récepteur FPRL1, est majoritairement dépendante de la protéine G héterotrimérique et qu'il n'y pas de signalisation transmise par les b-arrestines. Enfin, une analyse protéomique des complexes multi-protéiques bâtis autour du couple FPRL1/b-arrestine a été menée par la méthode TAP (Tandem Affinity Purification). Le complexe adaptateur AP3, homologue d'AP2 a été identifié comme partenaire des b-arrestines après stimulation du récepteur FPRL1Phagocytes are the first host defense line against pathogens. Their directed migration to infection sites and their microbicidal functions result from intracellular signaling cascades elicited by specific G protein-coupled receptors, the chemoattractant receptors. After ligand binding and G protein-mediated signaling events, receptors are phosphorylated and interact with the b-arrestins, which are scaffolding proteins participating to receptor internalization. Recent examples suggest that b-arrestins could also participate to signaling events. The work presented here is focused on the FPR family (Formyl Peptide Receptor) and specially the FPRL1 receptor (FPR-like 1), for which new agonists from bacterial origin or derived from human mitochondrial proteins have been identified. The phosphorylation of the FPRL1 receptor has been characterised and it has been shown that b-arrestins interact with this receptor and are essential to its internalization. Various approaches have concluded that the FPRL1-elicited rapid and transient activation of the MAP kinases ERK1/2 is mainly dependant on G protein and that there is no signaling events mediated by the b-arrestins. Finally, a proteomic study of the multiprotein complexes surrounded the couple FPRL1/b-arrestin has been conducted by the TAP method (Tandem Affinity Purification). The adaptor complex AP3, an homolog of AP2, has been identified as partner of b-arrestins after FPRL1 stimulation
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EFSTATHIOU, MOJAISKY IRENE. "Etude physiologique et genetique de clostridium acetobutylicum souche abkn8". Paris 6, 1987. http://www.theses.fr/1987PA066355.
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Khiari, Souad. "Problèmes inverses de points sources dans les modèles de transport dispersif de contaminants : identifiabilité et observabilité". Thesis, Compiègne, 2016. http://www.theses.fr/2016COMP2301.
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La recherche et les questions abordées dans cette thèse sont de type inverse : la reconstitution d'une source ponctuelle ou la complétion d'une donnée à la limite inconnue à l'extrémité du domaine dans les modèles paraboliques de transport de contaminants. La modélisation mathématique des problèmes de pollution des eaux fait intervenir deux traceurs, l'oxygène dissous (OD) et la demande biochimique en oxygène (DBO) qui est la quantité d'oxygène nécessaire à la biodégradation de la matière organique. En effet, au cours des procédés d'autoépuration, certaines bactéries aérobies jouent un rôle principal. Ces micro-organismes décomposent les matières organiques polluantes en utilisant l'oxygène dissous dans le milieu. Afin de compenser ces données manquantes, les champs, solutions du problème, sont observés directement ou indirectement. Les problèmes inverses qui en résultent sont quasi certainement mal-posés voire même sévèrement mal-posés pour la plupart. Dans cette thèse, nous proposons justement une analyse aussi poussée que possible sur la question de l'identifiabilité pour les deux problèmes inverses décrits ci-dessus. Nous avons démontré un résultat d'unicité pour des sources fixes dans le cas d'observations décalées. La réalité pour l'observation est nuancée et l'idéal n'est pas acquis ; des mesures directes sur la DBO sont difficiles à obtenir. En revanche collecter des données sur l'OD est possible en temps réel et avec un faible coût. La DBO est donc observée de façon indirecte, grâce au couplage dans le système de Streeter et Phelps, l'information passe de l'OD à la DBO. Pour ce problème aussi, nous avons produit un résultat d'unicité pour la reconstruction de la source ou puits ponctuel qui serait présent dans l'équation de transport sur l'OD. Nous avons ensuite examiné des questions annexes à l'identifiabilité telles que le degré d'instabilité des équations à résoudre. De ce type d'informations dépendent le comportement des méthodes numériques et des algorithmes de calcul à utiliserThe research and the questions approached on this thesis are inverse type : the reconstruction of point-wise source or the data completion problem in parabolic models of transport of contaminants. The mathematical modelling of the problems of water pollution includes two tracers, the dissolved oxygen (DO) and the biochemical demand in oxygen (BDO) which is the quantity of oxygen necessary for the biodegradation of organic matter. Indeed, during the biodegradation process, aerobic bacteria play a leading part. These micro-organisms decompose polluting organic matters by using the dissolved oxygen in the middle. To compensate these missing data, fields, solutions of the problem, are observed directly or indirectly. The resulting inverse problems are ill-posed. Their mathematical study rises big complications and their numerical treatment isn't easy. We demonstrated a uniqueness result for fixed sources in the case of moved observations. The reality for the observation is qualified and the ideal is not acquired; direct measures on the BOD are difficult to obtain. On the Other hand to collect data on the DO is possible in real time With a moderate cost. The BOD is thus observed in indirect way, thanks to the coupling in the system of Streeter and Phelps, the information passes from the DO to the BOD. For this problem, we produced a uniqueness result for the reconstruction of source. Then, we examined the degree of instability of the equation to be solved. The behaviour of numerical methods depend on this type of information
Książki na temat "MAB PURIFICATION"
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Ksenofontov, Boris, Aleksandr Lukanin i Evgeniy Pirogov. Chemical and physico-chemical methods of wastewater and man-made water treatment. ru: INFRA-M Academic Publishing LLC., 2022. http://dx.doi.org/10.12737/1863094.
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The monograph discusses issues related to chemical and physico-chemical methods of wastewater and man-made water treatment, including oxidation and sorption of pollutants, as well as their coagulation and flocculation and other processes of wastewater and man-made water treatment. At the same time, individual tasks are considered for the first time both in domestic and in the world practice of water treatment. First of all, this applies to the use of strong oxidizing agents in the practice of water purification, as well as neutralizing substances. In addition, the issues of intensification of chemical reactions occurring during wastewater treatment using various reagents are considered. At the same time, water treatment using ultraviolet light, electromagnetic fields, etc. is considered as an intensifying effect. As a result of the complex effect on the treated water, a high technological effect of purification is achieved.
For a wide range of readers, including researchers, university professors, graduate students, masters, bachelors and undergraduates.
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F, Stoltz J., i Rivat C, red. Biotechnologie des protéines du plasma--purification et utilisations cliniques et biologiques: Symposium international INSERM, Nancy, 17-19 mai 1988. Paris: Editions de l'Institut national de la santé et de la recherche médicale, 1989.
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Luft, VDI-Kommission Reinhaltung der, red. Biologische Abgasreinigung: Praktische Erfahrungen und neue Entwicklungen : Tagung, Köln, 23. und 24. Mai 1989. Düsseldorf: VDI Verlag, 1989.
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Christian, Hänsel, i Sächsische Akademie der Wissenschaften zu Leipzig., red. Gewässer und ihre Einzugsgebiete: Ökologische Ansätze zur Sanierung : ausgewählte Vorträge, gehalten auf der Klausurtagung der Sächsischen Akademie der Wissenschaften zu Leipzig, 11. bis 13. Mai 1994, Hydrobiologisches Laboratorium Neunzehnhain, Lengefeld/Erzgebirge. Berlin: Akademie Verlag, 1996.
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Padma-bdud-ʼdul. Bkaʼ gsaṅ zab chos mkhaʼ khyab raṅ grol las, skad cig gcig gis rdzogs saṅs rgyas paʼi myur lam saṅs rgyas kyi mtshan brjod ṅes ltuṅ druṅ nas ʼbyin pa: A formula for the purification of spiritual defects with other confessional texts. Darjeeling: Chopal Lama, 1985.
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Thunder, Robert S. Final Purification. iUniverse, Incorporated, 2007.
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Rao, John C. Centenary Meditation on a Quest for Purification Gone Mad: Gardone Lectures. Arouca Press, 2019.
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Frankfurter, David. The Construction of Evil and the Violence of Purification. Redaktorzy Michael Jerryson, Mark Juergensmeyer i Margo Kitts. Oxford University Press, 2013. http://dx.doi.org/10.1093/oxfordhb/9780199759996.013.0035.
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This chapter explores the construction of evil and the strategies of violence in purification. Prurient fascination and righteous revulsion both recreate and repel each other, developing an anxiety of confusion that has resulted in many circumstances in community efforts to cast the subject, the symbol, of that confusion. Erotic prurience into the nature and deeds of Evil may remain as a living genre for centuries without lending itself to societies as legitimation for purge. Dramaturgy and procession can contribute to brutal but cathartic narratives of saints and monsters, martyrs, and their persecutors, into the immediate festival lives of communities. Furthermore, brutality and atrocity are recurrent characteristics of any culture, often aggravated in situations of historical stress independent of religious systems.
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Umweltentlastung bei der Verbrennung: Tagung, Hamburg, 16. und 17. Mai 1988 = Combustion, pollution, reduction : new techniques in Europe. Düsseldorf: VDI-Verlag, 1988.
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Ozone-enhanced biofiltration for geosmin and MIB removal. Denver, CO: Awwa Research Foundation and American Water Works Association, 2005.
Znajdź pełny tekst źródłaCzęści książek na temat "MAB PURIFICATION"
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Valdés, R., B. Reyes, N. Ibarra, R. Hernández, M. González, S. Padilla, A. Tamayo i in. "Production of Mab CB.Hep-1 for the purification of rHBsAg at high scale". W Animal Cell Technology: Basic & Applied Aspects, 195–99. Dordrecht: Springer Netherlands, 2002. http://dx.doi.org/10.1007/978-94-017-0728-2_35.
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Pfahler, V., J. Adu-Gyamfi, A. Watzinger i F. Tamburini. "Modifications and Issues During Purification". W Oxygen Isotopes of Inorganic Phosphate in Environmental Samples, 45–49. Cham: Springer International Publishing, 2022. http://dx.doi.org/10.1007/978-3-030-97497-8_4.
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AbstractDepending on the extract, it is necessary to modify the purification protocol slightly. Each sample is different and despite a thorough testing of the purification protocol, issues might occur. The three modifications suggested include (1) adjustments in pH, (2) magnesium ammonium phosphate (MAP) precipitation and (3) reductions, prior to A1, of cations like iron (Fe), silica (Si) and calcium (Ca) which could cause interferences during the purification process. Some of the major issues often encountered are (1) no APM precipitation due to the presence of high carbonate concentrations, (2) the presence of high organic matter that requires additional steps in the protocol, (3) crystals not dissolving and (4) discoloration of solution.
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Guinó, Meritxell, i Yolanda R. de Miguel. "HR-MAS NMR Analysis of Compounds Attached to Polymer Supports". W Analysis and Purification Methods in Combinatorial Chemistry, 71–86. Hoboken, NJ, USA: John Wiley & Sons, Inc., 2004. http://dx.doi.org/10.1002/0471531979.ch4.
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Bohman, Christina, i Staffan Eriksson. "Purification and Properties of Human Deoxycytidine Kinase". W Purine and Pyrimidine Metabolism in Man V, 311–14. New York, NY: Springer US, 1986. http://dx.doi.org/10.1007/978-1-4684-1248-2_49.
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Hards, R. G., S. L. Graw i D. Patterson. "Purification of Mammalian Glycinamide Ribonucleotide (GAR) Synthetase". W Purine and Pyrimidine Metabolism in Man V, 315–19. New York, NY: Springer US, 1986. http://dx.doi.org/10.1007/978-1-4684-1248-2_50.
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Edmondson, Dale E. "Purification of MAO A and MAO B from Mammalian Tissue Sources". W Methods in Molecular Biology, 1–10. New York, NY: Springer US, 2022. http://dx.doi.org/10.1007/978-1-0716-2643-6_1.
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Lindmark Månsson, H., i B. Åkesson. "Purification and Enzyme-Linked Immunoassay of Bovine Extracellular Glutathione Peroxidase". W Trace Elements in Man and Animals 10, 893–94. New York, NY: Springer US, 2002. http://dx.doi.org/10.1007/0-306-47466-2_281.
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Deagen, J. T., D. J. Broderick i P. D. Whanger. "Purification and Properties of the Human Plasma Selenoenzyme, Glutathione Peroxidase". W Trace Elements in Man and Animals 6, 339–40. Boston, MA: Springer US, 1988. http://dx.doi.org/10.1007/978-1-4613-0723-5_110.
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Edmondson, Dale E. "Purification of Recombinant Eukaryotic MAO A and MAO B Utilizing the Pichia pastoris Expression System". W Methods in Molecular Biology, 11–22. New York, NY: Springer US, 2022. http://dx.doi.org/10.1007/978-1-0716-2643-6_2.
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Penna, Thereza Christina Vessoni, Marina Ishii, Adalberto Pessoa Junior, Laura Oliveira de Nascimento, Luciana Cambricoli de Souza i Olivia Cholewa. "Evaluation of Recombinant Green Fluorescent Protein, Under Various Culture Conditions and Purification with HiTrap Hydrophobic Interaction Chromatography Resins". W Proceedings of the Twenty-Fifth Symposium on Biotechnology for Fuels and Chemicals Held May 4–7, 2003, in Breckenridge, CO, 453–68. Totowa, NJ: Humana Press, 2004. http://dx.doi.org/10.1007/978-1-59259-837-3_39.
Pełny tekst źródłaStreszczenia konferencji na temat "MAB PURIFICATION"
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Smith, K. J. "INFUSION OF MONOCLONAL ANTIBODY IMMUNOAFFINITY PURIFIED FACTOR IX IN RABBITS: COMPARISON WITH COMMERCIAL CONCENTRATES". W XIth International Congress on Thrombosis and Haemostasis. Schattauer GmbH, 1987. http://dx.doi.org/10.1055/s-0038-1644066.
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Commercial concentrates (CC) of vitamin K dependent coagulation factors may cause thrombosis or coagulation factor consumption while more highly purified (17 U/mg) factor IX (IX) concentrates do not seem to be thrombogenic (Menache et al, Blood 64:1220, 1984). Monoclonal antibody (MAb) immunoaffinity purified IX of high specific activity from CC or recombinant factor IX sources may also improve therapy. In this report, 400 mg of Affigel-10 linked A-7 MAb was used to bind factor IX in the presence of metal ions (20 mM MgCl2). Elution of IX was with 20 mM EDTA. Thrombogenicity of CC and IX prepared from CC by MAb immunoaffinity was tested. A CC which was thrombogenic in the stasis thrombosis assay (CC #1) produced large thrombi at doses of 50, 50, and 100 U/kg while none were seen with the IX produced from this CC at doses of 106 and 234 U/kg. A heparin treated CC (CC #2) which was not thrombogenic in the stasis-thrombosis assay at doses of 100 U/kg was infused in 4 rabbits at 100 U/kg and platelets, fibrinogen, AT-III antigen, and factors IX, V, and VIII were monitored for 5 hours post-infusion. Immunoaffinity IX from this CC was infused in 4 rabbits at 214-243 U/kg for comparison. Mean platelet count decrease was 20% in CC group and 8% for the IX group. Mean factor V and VIII decreased 26 and 36% respectively with CC while no decrease was seen in the IX rabbits (p < .05). Fibrinogen values and AT-III did not differ for IX or CC groups. Mean factor IX activity at 1 hour increased 1.6 fold for CC and 3 fold for IX. Yields for IX purification were 80 and 85%. Clotting activity was 143 and 101 U/mg and antigen was approximately 200 U/mg. Purification was over 90 fold by MAb immunoaf f inity. There was no detectable factor II, VII or X activity in MAb purified IX. Non-activated PTT was greater than 200 seconds for CC #1 and 158 seconds for CC #2. Column capacity was at least 150 mg. These results demonstrate that factor IX is not the thrombogenic component of some CC. Also, IX prepared by MAb immunoaffinity may have therapeutic advantage for patients at risk for thrombosis and adverse effects of contaminating proteins in commercial concentrates.
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Marcovina, S., R. Coppola, M. P. Protti, C. Gelfi i P. M. Mannucci. "EDTA-DEPENDENT MONOCLONAL ANTIBODIES TO HUMAN PROTEIN S". W XIth International Congress on Thrombosis and Haemostasis. Schattauer GmbH, 1987. http://dx.doi.org/10.1055/s-0038-1644294.
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Splenocytes from a Balb/c mouse immunized with purified human protein S (PS) were fused with murine hybridoma cell line SP2/0-Agl4 and cultured in Iscove's medium without addition of fetal bovine serum. Hybrid supernatants were screened for the presence of specific antibodies by solid-phase ELISA and cloned by the limiting dilution technique. Pour clones, named S2, S3, S8, and S10, were selected, recloned several times, and injected intraperitoneally into Balb/c mice for the production of antibody-rich ascitic fluid. The monoclonal antibodies (Mabs), all of IgGl subclass with k light chain, were purified from ascitic fluid by Protein-A chromatography. The specificity of Mabs was controlled by the immunoblotting technique: the Mabs appeared to react only with two plasma proteins, one with a MW of about 70.000 dal tons comigrating with purified PS, and the other with a MW >300.000 da that is likely to be the C4b-binding protein-PS complex. No interaction has been observed with PS-depleted plasma. As tested by a fluid phase radio immunoassay, the binding of Mabs to PS was significantly higher in the presence of EDTA while was totally inhibited in the presence of Ca2+. Scatchard plot analysis of the binding between 125I-PS and Mabs showed that the binding affinity of the antibodies ranged from 108 to 109 1/mol. Each EDTA-dependent Mab was then immobilized on Sepharose 4B-CNBr and used to purify PS from barium precipitation of citrated plasma. The fraction eluted with 2 mmol of CaCl2 from the immunoadsorbent appeared to contain only two proteins when stained with Cocmassie Blue. By immuno blotting, all Mabs reacted with both proteins, one comigrating with purified PS and the other with a MW >300.000. Essentially homogeneous PS, free from the high MW component, was obtained when the barium citrate adsorbate was applied to a DEAE-Sephadex column and the protein peack containing the balk of PS was sussequently applied to the immunoadsorbent and eluted with 2 mmol CaCl2.In summary, we have described an unusual set of EDTA-dependent monoclonal antibodies to PS and their use for the purification of homogeneous protein S from human plasma.
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Wang, Yutian, Songnian Fu, Ming Tang, Chi Zhang, Xiahui Tang, Jian Kong i Luming Zhao. "Nonlinear Fourier Transform Enabled Multiple Pulses Purification for Soliton Communication". W Optical Fiber Communication Conference. Washington, D.C.: OSA, 2021. http://dx.doi.org/10.1364/ofc.2021.m5b.6.
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Freeman, Eric, Lisa Mauck Weiland i Ryan Soncini. "Water Purification Through Selective Transport". W ASME 2011 Conference on Smart Materials, Adaptive Structures and Intelligent Systems. ASMEDC, 2011. http://dx.doi.org/10.1115/smasis2011-5062.
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Accumulation of inorganic nitrates and phosphates in regions such as the Mississippi river basin has resulted in catastrophic growth of algal blooms. These algal blooms deplete the surrounding oxygen and asphyxiate nearby aquatic life, resulting in large regions incapable of sustaining life. Using biomimicry principles to design a tailored active material to selectively transport these pollutants may offer a strategy to restore these dead zones to health. Theoretically a combination of selected protein transporters may be employed to create a selective sponge to reclaim these nitrates and phosphates. Presented is a feasibility study of various configurations of transporters, and a unique solution for restoring the aquatic ecosystem back to health.
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Duff, William S., i David Hodgson. "Solar Water Purification by Pasteurization". W ASME 2003 International Solar Energy Conference. ASMEDC, 2003. http://dx.doi.org/10.1115/isec2003-44214.
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A new passive solar water pasteurization system has been designed, built and tested. The system contains no valves and regulates flow based on the density difference between two columns of water. The new system eliminates boiling problems encountered in previous designs. Boiling is undesirable because it may contaminate treated water. The system has produced over 100 liters per day of treated water with a collector area of 0.45m2. Work is ongoing to develop a theoretical understanding of system behavior, to analytically model it and to further improve system performance.
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Stevens, R. P., V. Solodushko, V. V. Pastukh, T. Stevens, R. Honkanen, M. Swingle i J. Y. Lee. "Expression and Purification of the Human Phosphatase PPM1B". W American Thoracic Society 2023 International Conference, May 19-24, 2023 - Washington, DC. American Thoracic Society, 2023. http://dx.doi.org/10.1164/ajrccm-conference.2023.207.1_meetingabstracts.a2020.
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Wu, Wei, Haijun Hu, Yuhai Lv, Weiguo Zeng, Xiufeng Li, Ke Song i Xinshuang Du. "An Alternative Corrosion Risk Assessment Method for Industrial Pipelines in Natural Gas Purification Plant". W ASME 2020 Pressure Vessels & Piping Conference. American Society of Mechanical Engineers, 2020. http://dx.doi.org/10.1115/pvp2020-21376.
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Abstract Natural gas is a kind of high-quality and low-carbon energy and its demand is increasing rapidly year by year. The natural gas produced from the gas well contains some corrosive impurities like CO2, H2S, and H2O. Therefore, the gas needs to be purified in purification plants before it can be used. In the process of purification, the impurities in the natural gas will cause corrosion to the purification equipment, especially to the industrial pipeline, which will lead to perforation and leakage of the pipeline, and even cause poisoning, fire or explosion, and other serious safety accidents. In order to ensure the safe operation of the industrial pipeline in the purification plant, it is necessary to carry out effective inspection and maintenance measures for the pipelines. The traditional regular inspection method just inspects a certain proportion of the pipelines which are selected by on-site inspection personnel. But limited by technical level and work experience of the inspection personnel, some high-risk pipelines may be neglected. Therefore, some plants try to adopt risk-based inspection (RBI) to carry out a risk assessment and conduct a comprehensive inspection on high-risk pipelines, so as to improve inspection efficiency and reduce inspection cost. However, in practice, it was found that the pipeline risk levels are not in accordance with the actual inspection results. The reason is that the RBI method developed in the background of oil refining and chemical industry is not suitable for natural gas purification plants. In order to solve the problems, this paper analyzed the relationship between the influencing factors and pipeline corrosion behavior. The influencing factors include the impurity contents (CO2, H2S, H2O, and chlorides), pipeline materials (carbon steel and austenitic stainless steel), and service conditions (operating temperature, operating pressure and flow velocity). Meanwhile, the plant management status and maintenance factors were also considered as the influencing factors to pipelines. According to the analysis, a corrosion risk assessment method for the pipelines in the natural gas purification plant was developed based on fault tree and scoring method. Finally, the method was applied to the pipelines in purification plant to verify the accuracy of this method.
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Tamburello, David, Bruce Hardy i Martin Sulic. "Multi-Component Separation and Purification of Natural Gas". W ASME 2018 Power Conference collocated with the ASME 2018 12th International Conference on Energy Sustainability and the ASME 2018 Nuclear Forum. American Society of Mechanical Engineers, 2018. http://dx.doi.org/10.1115/power2018-7537.
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Over the past decade, several technical developments (such as hydraulic fracturing) have led to an exponential increase in discovering new domestic natural gas reserves. Raw natural gas composition can vary substantially from source to source. Typically, methane accounts for 75% to 95% of the total gas, with the rest of the gas containing ethane, propane, butane, other higher hydrocarbons, and impurities, with the most common including H2O, CO2, N2, and H2S. All natural gas requires some treatment, if only to remove H2O; however, the composition of natural gas delivered to the commercial pipeline grids is tightly controlled. Sub-quality natural gas reserves, which are defined as fields containing more than 2% CO2, 4% N2, or 4 ppm H2S, make up nearly half of the world’s natural gas volume. The development of sub-quality, remote, and unconventional fields (i.e. landfill gas) can present new challenges to gas separation and purification methods. Adsorbent technologies, such as the use of activated carbons, zeolites, or metal-organic frameworks (MOFs), may hold the key to more efficient and economically viable separation methods. This work proposes to prove the applicability of the multi-component potential theory of adsorption (MPTA) to a real world natural gas adsorbent system to properly characterize the adsorbent’s selectivity for an individual gas component using only the single component isotherms. Thus, the real-world gas separation/purification application of a specific adsorbent for a given gas stream can be obtained simply and effectively without the need for large experimental efforts or costly system modifications until after an initial computational screening of perspective materials has been completed. While the current research effort will use natural gas, which is the world’s largest industrial gas separations application, to validate the MPTA, the tools gained through this effort can be applied to other gas separation effort.
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Shen, Kui, Lishi Xie, Grace R. Shen, Steven Dudek i Joe G. N. Garcia. "Expression And Purification Of The N-Terminal Segments Of NmMLCK For Ligand-Binding Analysis". W American Thoracic Society 2012 International Conference, May 18-23, 2012 • San Francisco, California. American Thoracic Society, 2012. http://dx.doi.org/10.1164/ajrccm-conference.2012.185.1_meetingabstracts.a1868.
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Gordon, Stuart, Bonnie Sloane, Phil Cavanugh, Barbara Cross, Kenneth Honn i Mohanathasan Chelladurai. "PURIFICATION AND CHARACTERIZATION OF TWO PROCOAGULANTS FROM WALKER 256 CARCINOSARCOMA TUMORS",. W XIth International Congress on Thrombosis and Haemostasis. Schattauer GmbH, 1987. http://dx.doi.org/10.1055/s-0038-1643666.
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Activation of the coagulation system bytumor cells may play an important role in tumor growth and metastases. Becauseprocoagulant activities have been identified in different tumor cells by different investigators, effective comparison of these activities has been difficult. Therefore, we purified and characterized two different procoagulant proteins from the same Walker 256 tumors. The first procoagulant activity/platelet aggregating activity (PCA/PAA) was purified from a 1% CHAPS detergent extract oftumor homogenate followed by (NH4)2SO4 fractionation, anion exchange and hydrophobic chromatography. The protein had a molecular weight of 58,000, required phospholipid and an intact coagulation pathway from factor X through fibrinogen for activity, but did not require factors VII or IX forits procoagulant activity. The procoagulant activity was not inhibited by 5mMphenyl-methyl sulfonyl fluoride, iodoacetamide or phenanthroline; there was noevidence of proteinase activity. The PAA was due to thrombin generation during coagulation. The second procoagulant,cancer procoagulant (CP), was extracted from tumors in barbital buffer (pH 7.4) without detergent, purified by immunoaffinity (using a polyclonal goat antibody to CP from V2 carcinoma) and mercurial-benzoate affinity chromatography. CP had a molecular weight of 68,000, an isoelectric point of 4.8 and initiated coagulation by directly activating factor X in the coagulation system. CP was inhibited by Hg++ and iodoacetamide, cysteine proteinase inhibitors. The purified CP formed an immunodiffusion precipitin band against the polyclonal anti-CP goat antibody. Thus, thepurified CP had the same physicochemical, enzymatic and immunologic propertiesas CP from rabbit V2 carcinoma. Neither procoagulant had the properties of tissue factor. These results suggest that there aretwo distinct procoagulant activities inWalker 256 and that both may contributeto the coagulation abnormalities that are associated with tumor growthand metastases.
Raporty organizacyjne na temat "MAB PURIFICATION"
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Lemar, Homer J., Georgitis Jr., McDermott William J. i Michael T. Effect of Prolonged Administration of Iodine Containing Water Purification Tablets in Man. Fort Belvoir, VA: Defense Technical Information Center, kwiecień 1993. http://dx.doi.org/10.21236/ada267047.
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Rafaeli, Ada, Russell Jurenka i Daniel Segal. Isolation, Purification and Sequence Determination of Pheromonotropic-Receptors. United States Department of Agriculture, lipiec 2003. http://dx.doi.org/10.32747/2003.7695850.bard.
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Moths constitute a major group of pest insects in agriculture. Pheromone blends are utilised by a variety of moth species to attract conspecific mates, which is under circadian control by the neurohormone, PBAN (pheromone-biosynthesis-activating neuropeptide). Our working hypothesis was that, since the emission of sex-pheromone is necessary to attract a mate, then failure to produce and emit pheromone is a potential strategy for manipulating adult moth behavior. The project aimed at identifying, characterising and determining the sequence of specific receptors responsible for the interaction with pheromonotropic neuropeptide/s using two related moth species: Helicoverpa armigera and H. lea as model insects. We established specific binding to a membrane protein estimated at 50 kDa in mature adult females using a photoaffinity-biotin probe for PBAN. We showed that JH is required for the up-regulation of this putative receptor protein. In vitro studies established that the binding initiates a cascade of second messengers including channel opening for calcium ions and intracellular cAMP production. Pharmacological studies (using sodium fluoride) established that the receptor is coupled to a G-protein, that is, the pheromone-biosynthesis-activating neuropeptide receptor (PBAN-R) belongs to the family of G protein-coupled receptor (GPCR)'s. We showed that PBAN-like peptides are present in Drosophila melanogaster based on bioassay and immunocytochemical data. Using the annotated genome of D. melanogaster to search for a GPCR, we found that some were similar to neuromedin U- receptors of vertebrates, which contain a similar C-terminal ending as PBAN. We established that neuromedin U does indeed induce pheromone biosynthesis and cAMP production. Using a PCR based cloning strategy and mRNA isolated from pheromone glands of H. zea, we successfully identified a gene encoding a GPCR from pheromone glands. The full-length PBAN-R was subsequently cloned and expressed in Sf9 insect cells and was shown to mobilize calcium in response to PBAN in a dose-dependent manner. The successful progress in the identification of a gene, encoding a GPCR for the neurohormone, PBAN, provides a basis for the design of a novel battery of compounds that will specifically antagonize pheromone production. Furthermore, since PBAN belongs to a family of insect neuropeptides with more than one function in different life stages, this rationale may be extended to other physiological key-regulatory processes in different insects.
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Chefetz, Benny, Baoshan Xing, Leor Eshed-Williams, Tamara Polubesova i Jason Unrine. DOM affected behavior of manufactured nanoparticles in soil-plant system. United States Department of Agriculture, styczeń 2016. http://dx.doi.org/10.32747/2016.7604286.bard.
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The overall goal of this project was to elucidate the role of dissolved organic matter (DOM) in soil retention, bioavailability and plant uptake of silver and cerium oxide NPs. The environmental risks of manufactured nanoparticles (NPs) are attracting increasing attention from both industrial and scientific communities. These NPs have shown to be taken-up, translocated and bio- accumulated in plant edible parts. However, very little is known about the behavior of NPs in soil-plant system as affected by dissolved organic matter (DOM). Thus DOM effect on NPs behavior is critical to assessing the environmental fate and risks related to NP exposure. Carbon-based nanomaterials embedded with metal NPs demonstrate a great potential to serve as catalyst and disinfectors. Hence, synthesis of novel carbon-based nanocomposites and testing them in the environmentally relevant conditions (particularly in the DOM presence) is important for their implementation in water purification. Sorption of DOM on Ag-Ag₂S NPs, CeO₂ NPs and synthesized Ag-Fe₃O₄-carbon nanotubebifunctional composite has been studied. High DOM concentration (50mg/L) decreased the adsorptive and catalytic efficiencies of all synthesized NPs. Recyclable Ag-Fe₃O₄-carbon nanotube composite exhibited excellent catalytic and anti-bacterial action, providing complete reduction of common pollutants and inactivating gram-negative and gram-positive bacteria at environmentally relevant DOM concentrations (5-10 mg/L). Our composite material may be suitable for water purification ranging from natural to the industrial waste effluents. We also examined the role of maize (Zeamays L.)-derived root exudates (a form of DOM) and their components on the aggregation and dissolution of CuONPs in the rhizosphere. Root exudates (RE) significantly inhibited the aggregation of CuONPs regardless of ionic strength and electrolyte type. With RE, the critical coagulation concentration of CuONPs in NaCl shifted from 30 to 125 mM and the value in CaCl₂ shifted from 4 to 20 mM. This inhibition was correlated with molecular weight (MW) of RE fractions. Higher MW fraction (> 10 kDa) reduced the aggregation most. RE also significantly promoted the dissolution of CuONPs and lower MW fraction (< 3 kDa) RE mainly contributed to this process. Also, Cu accumulation in plant root tissues was significantly enhanced by RE. This study provides useful insights into the interactions between RE and CuONPs, which is of significance for the safe use of CuONPs-based antimicrobial products in agricultural production. Wheat root exudates (RE) had high reducing ability to convert Ag+ to nAg under light exposure. Photo-induced reduction of Ag+ to nAg in pristine RE was mainly attributed to the 0-3 kDa fraction. Quantification of the silver species change over time suggested that Cl⁻ played an important role in photoconversion of Ag+ to nAg through the formation and redox cycling of photoreactiveAgCl. Potential electron donors for the photoreduction of Ag+ were identified to be reducing sugars and organic acids of low MW. Meanwhile, the stabilization of the formed particles was controlled by both low (0-3 kDa) and high (>3 kDa) MW molecules. This work provides new information for the formation mechanism of metal nanoparticles mediated by RE, which may further our understanding of the biogeochemical cycling and toxicity of heavy metal ions in agricultural and environmental systems. Copper sulfide nanoparticles (CuSNPs) at 1:1 and 1:4 ratios of Cu and S were synthesized, and their respective antifungal efficacy was evaluated against the pathogenic activity of Gibberellafujikuroi(Bakanae disease) in rice (Oryza sativa). In a 2-d in vitro study, CuS decreased G. fujikuroiColony- Forming Units (CFU) compared to controls. In a greenhouse study, treating with CuSNPs at 50 mg/L at the seed stage significantly decreased disease incidence on rice while the commercial Cu-based pesticide Kocide 3000 had no impact on disease. Foliar-applied CuONPs and CuS (1:1) NPs decreased disease incidence by 30.0 and 32.5%, respectively, which outperformed CuS (1:4) NPs (15%) and Kocide 3000 (12.5%). CuS (1:4) NPs also modulated the shoot salicylic acid (SA) and Jasmonic acid (JA) production to enhance the plant defense mechanisms against G. fujikuroiinfection. These results are useful for improving the delivery efficiency of agrichemicals via nano-enabled strategies while minimizing their environmental impact, and advance our understanding of the defense mechanisms triggered by the NPs presence in plants.
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Barefoot, Susan, Benjamin Juven, Thomas Hughes, Avraham Lalazar, A. B. Bodine, Yitzhak Ittah i Bonita Glatz. Characterization of Bacteriocins Produced by Food Bioprocessing Propionobacteria. United States Department of Agriculture, sierpień 1992. http://dx.doi.org/10.32747/1992.7561061.bard.
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Objectives were to further characterize activity spectra of dairy propionibacteria bacteriocins, jenseniin G and propionicin PLG-1, purify them, examine the role of cell walls in resistance, examine their interactions with cytoplasmic membrane, explain producer immunity, and clone the responsible genes. Inhibitory spectra of both bacteriocins were further characterized. Propionicin was most effective in controlling Gram-positive, rather than Gram-negative organisms; it controlled growth of sensitive cells both in a culture medium and a model food system. Jenseniin inhibited yogurt cultures and may help prevent yogurt over-acidification. Both were active against botulinal spores; jenseniin was sporostatic; propionicin was sporicidal. Jenseniin was produced in broth culture, was stable to pH and temperature extremes, and was purified. Its molecular mass (3649 Da) and partial amino acid composition (74%) were determined. A blocked jenseniin N-terminus prevented sequencing. Methods to produce propionicin in liquid culture were improved, and large scale culture protocols to yield high titers were developed. Methods to detect and quantify propionicin activity were optimized and standardized. Stability of partially purified propionicin was demonstrated and an improved purification scheme was developed. Purified propionicin had a 9328-Da molecular mass, contained 99 amino acids, and was significantly hydrophobic; ten N-terminal amino acids were identified. Propionicin and Jenseniin interacted with cytoplasmic membranes; resistance of insensitive species was cell wall-related. Propionicin and jenseniin acted similarly; their mode of action appeared to differ from nisin. Spontaneous jenseniin-resistant mutants were resistant to propionicin but nisin-sensitive. The basis for producer immunity was not resolved. Although bacteriocin genes were not cloned, a jenseniin producer DNA clone bank and three possible vectors for cloning genes in propionibacteria were constructed. In addition, transposon Tn916 was conjugatively transferred to the propionicin producer from chromosomal and plasmid locations at transfer frequencies high enough to permit use of Tn916 for insertional mutagenesis or targeting genes in propionibacteria. The results provide information about the bacteriocins that further supports their usefulness as adjuncts to increase food safety and/or quality.
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Nelson, Nathan, i Charles F. Yocum. Structure, Function and Utilization of Plant Photosynthetic Reaction Centers. United States Department of Agriculture, wrzesień 2012. http://dx.doi.org/10.32747/2012.7699846.bard.
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Light capturing and energy conversion by PSI is one of the most fundamental processes in nature. In the heart of these adaptations stand PSI, PSII and their light harvesting antenna complexes. The main goal of this grant proposal was to obtain by X-ray crystallography information on the structure of plant photosystem I (PSI) and photosystem II (PSII) supercomplexes. We achieved several milestones along this line but as yet, like several strong laboratories around the world, we have no crystal structure of plant PSII. We have redesigned the purification and crystallization procedures and recently solved the crystal structure of the PSI supercomplex at 3.3 Å resolution. Even though this advance in resolution appears to be relatively small, we obtained a significantly improved model of the supercomplex. The work was published in J. Biol. Chem. (Amunts et al., 2010). The improved electron density map yielded identification and tracing of the PsaK subunit. The location of an additional 10 ß-carotenes, as well as 5 chlorophylls and several loop regions that were previously uninterruptable have been modeled. This represents the most complete plant PSI structure obtained thus far, revealing the locations of and interactions among 17 protein subunits and 193 non-covalently bound photochemical cofactors. We have continued extensive experimental efforts to improve the structure of plant PSI and to obtain PSII preparation amenable to crystallization. Most of our efforts were devoted to obtain well-defined subcomplexes of plant PSII preparations that are amenable to crystallization. We studied the apparent paradox of the high sensitivity of oxygen evolution of isolated thylakoids while BBY particles exhibit remarkable resilience to the same treatment. The integrity of the photosystem II (PSII) extrinsic protein complement as well as calcium effects arise from the Ca2+ atom associated with the site of photosynthetic water oxidation were investigated. This work provides deeper insights into the interaction of PsbO with PSII. Sight-directed mutagenesis indicated the location of critical sites involved in the stability of the water oxidation reaction. When combined with previous results, the data lead to a more detailed model for PsbO binding in eukaryotic PSII.
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Husson, Scott M., Viatcheslav Freger i Moshe Herzberg. Antimicrobial and fouling-resistant membranes for treatment of agricultural and municipal wastewater. United States Department of Agriculture, styczeń 2013. http://dx.doi.org/10.32747/2013.7598151.bard.
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This research project introduced a novel membrane coating strategy to combat biofouling, which is a major problem for the membrane-based treatment of agricultural and municipal wastewaters. The novelty of the strategy is that the membrane coatings have the unique ability to switch reversibly between passive (antifouling) and active (antimicrobial) fouling control mechanisms. This dual-mode approach differs fundamentally from other coating strategies that rely solely on one mode of fouling control. The research project had two complementary objectives: (1) preparation, characterization, and testing of dual-mode polymer nanolayers on planar surfaces and (2) evaluation of these nanolayers as membrane modifiers. The first objective was designed to provide a fundamental understanding of how polymer nanolayer chemistry and structure affect bacterial deposition and to demonstrate the reversibility of chemical switching. The second objective, which focused on membrane development, characterization, and testing, was designed to demonstrate methods for the production of water treatment membranes that couple passive and active biofouling control mechanisms. Both objectives were attained through synergistic collaboration among the three research groups. Using planar silicon and glass surfaces, we demonstrated using infrared spectroscopy that this new polymer coating can switch reversibly between the anti-fouling, zwitterion mode and an anti-microbial, quaternary amine mode. We showed that switching could be done more than 50 times without loss of activity and that the kinetics for switching from a low fouling zwitterion surface to an antimicrobial quaternary amine surface is practical for use. While a low pH was required for switching in the original polymer, we illustrated that by slightly altering the chemistry, it is possible to adjust the pH at which the switching occurs. A method was developed for applying the new zwitterionic surface chemistry onto polyethersulfone (PES) ultrafiltration membranes. Bacteria deposition studies showed that the new chemistry performed better than other common anti-fouling chemistries. Biofilm studies showed that PESultrafiltration membranes coated with the new chemistry accumulated half the biomass volume as unmodified membranes. Biofilm studies also showed that PES membranes coated with the new chemistry in the anti-microbial mode attained higher biofilm mortality than PES membranes coated with a common, non-switchablezwitterionic polymer. Results from our research are expected to improve membrane performance for the purification of wastewaters prior to use in irrigation. Since reduction in flux due to biofouling is one of the largest costs associated with membrane processes in water treatment, using dual-mode nanolayer coatings that switch between passive and active control of biofouling and enable detachment of attached biofoulants would have significant economic and societal impacts. Specifically, this research program developed and tested advanced ultrafiltration membranes for the treatment of wastewaters. Such membranes could find use in membrane bioreactors treating municipal wastewater, a slightly upgraded version of what presently is used in Israel for irrigation. They also may find use for pretreatment of agricultural wastewaters, e.g., rendering facility wastewater, prior to reverse osmosis for desalination. The need to desalinate such impaired waters water for unlimited agricultural use is likely in the near future.
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Chamovitz, Daniel A., i Albrecht G. Von Arnim. eIF3 Complexes and the eIF3e Subunit in Arabidopsis Development and Translation Initiation. United States Department of Agriculture, wrzesień 2009. http://dx.doi.org/10.32747/2009.7696545.bard.
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The original working hypothesis of our proposal was that The “e” subunit of eIF3 has multiple functions from both within the nucleus and in the cytoplasm. Within this model, we further hypothesized that the “e” subunit of eIF3 functions in translation as a repressor. We proposed to test these hypotheses along the following specific aims: 1) Determine the subcellular localization of the interaction between eIF3e and other eIF3 subunits, or the COP9 signalosome. 2) Elucidate the biological significance of the varied subcellular localizations of eIF3e through generating Arabidopsis eIF3e alleles with altered subcellular localization. 3.) Purify different eIF3e complexes by tandem affinity purification (TAP). 4) Study the role of eIF3e in translational repression using both in vitro and in planta assays. eIF3 is an evolutionarily ancient and essential component of the translational apparatus in both the plant and animal kingdoms. eIF3 is the largest, and in some ways the most mysterious, of the translation factors. It is a multi-subunit protein complex that has a structural/scaffolding role in translation initiation. However, despite years of study, only recently have differential roles for eIF3 in the developmental regulation of translation been experimentally grounded. Furthermore, the roles of individual eIF3 subunits are not clear, and indeed some, such as the “e” subunit may have roles independent of translation initiation. The original three goals of the proposal were technically hampered by a finding that became evident during the course of the research – Any attempt to make transgenic plants that expressed eIF3e wt or eIF3e variants resulted in seedling lethality or seed inviability. That is, it was impossible to regenerate any transgenic plants that expressed eIF3e. We did manage to generate plants that expressed an inducible form of eIF3e. This also eventually led to lethality, but was very useful in elucidating the 4th goal of the research (Yahalom et al., 2008), where we showed, for the first time in any organism, that eIF3e has a repressory role in translation. In attempt to solve the expression problems, we also tried expression from the native promoter, and as such analyzed this promoter in transgenic plants (Epel, 2008). As such, several additional avenues were pursued. 1) We investigated protein-protein interactions of eIF3e (Paz-Aviram et al., 2008). 2) The results from goal #4 led to a novel hypothesis that the interaction of eIF3e and the CSN meets at the control of protein degradation of nascent proteins. In other words, that the block in translation seen in csn and eIF3e-overexpressing plants (Yahalom et al., 2008) leads to proteasome stress. Indeed we showed that both over expression of eIF3e and the csn mutants lead to the unfolded protein response. 3) We further investigated the role of an additional eIF3 subunit, eIF3h, in transalational regulation in the apical meristem (Zhou et al., 2009). Epel, A. (2008). Characterization of eIF3e in the model plant Arabidopsis thaliana. In Plant Sciences (Tel Aviv, Tel Aviv University). Paz-Aviram, T., Yahalom, A., and Chamovitz, D.A. (2008). Arabidopsis eIF3e interacts with subunits of the ribosome, Cop9 signalosome and proteasome. Plant Signaling and Behaviour 3, 409-411. Yahalom, A., Kim, T.H., Roy, B., Singer, R., von Arnim, A.G., and Chamovitz, D.A. (2008). Arabidopsis eIF3e is regulated by the COP9 signalosome and has an impact on development and protein translation. Plant J 53, 300-311. Zhou, F., Dunlap, J.R., and von Arnim, A.G. The translation initiation factor subunit eIF3h is .1 involved in Arabidopsis shoot apical meristem maintenance and auxin response. (submitted to Development).
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Koven, William, Gordon Grau, Benny Ron i Tetsuya Hirano. Improving fry quality, survival and growth in commercially farmed fish by dietary stimulation of thyroid hormone production in premetamorphosing larvae. United States Department of Agriculture, 2004. http://dx.doi.org/10.32747/2004.7695856.bard.
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There is a direct correlation between successful metamorphosis from larvae to post-larvae and the quality of the resultant juveniles or fry. Juvenile quality, in turn, is a major factor influencing fish production level and market price. However, following the profound morphological and physiological changes occurring during metamorphosis, the emerging juveniles in some species characteristically demonstrate heterotrophic growth, poor pigmentation, cannibalism and generally poor survival. The white grouper (Epinephelus aeneus) in Israel and the Pacific threadfin (Polydactylussexfilis) in Hawaii are two promising candidates for mariculture that have high market value but a natural fishery that has sharply declined in recent years. Unfortunately, their potential for culture is severely hampered by variable metamorphic success limiting their production. The main objective was to compare the efficacy and economic viability of dietary or environmental iodine on metamorphic success and juvenile quality in the white grouper and the pink snapper which would lead to improved commercial rearing protocols and increased production of these species both in Israel and the US. The Hawaii Institute of Marine Biology encountered problems with the availability of pink snapper brood stock and larvae and changed to Pacific threadfin or moi which is rapidly becoming a premier aquaculture species in Hawaii and throughout the Indo-Pacific. The white grouper brood stock at the National Center for Mariculture was lost as a result of a viral outbreak following the sudden breakdown of the ozone purification system. In addition, the NCM suffered a devastating fire in the fall of 2007 that completely destroyed the hatchery and laboratory facilities although the BARD project samples were saved. Nevertheless, by studying alternate species a number of valuable findings and conclusions that can contribute to improved metamorphosis in commercially valuable marine species resulted from this collaborative effort. The Israeli group found that exposing white grouper larvae to external TH levels synchronized and increased the rate of metamorphosis. This suggested that sub-optimal synthesis of TH may be a major factor causing size heterogeneity in the larval population and high mortality through cannibalism by their larger more metamorphosed cohorts. Two protocols were developed to enrich the larvae with higher levels of the TH precursor, iodine; feeding iodine enriched Artemia or increasing the level of seawater iodine the larvae are exposed to. Results of accumulated iodine in gilthead seabream larvae indicated that the absorption of iodine from the water is markedly more efficient than feeding iodine enriched Artemia nauplii. Samples for TH, which will be analyzed shortly, will be able to determine if another dietary factor is lacking to effectively utilize surplus tissue iodine for TH synthesis. Moreover, these samples will also clarify which approach to enriching larvae with iodine, through the live food or exposure to iodine enriched seawater is the most efficient and cost effective. The American group found that moi larvae reared in ocean water, which possessed substantially higher iodine levels than those found in seawater well water, grew significantly larger, and showed increased survival compared with well water reared larvae. Larvae reared in ocean water also progressed more rapidly through developmental stages than those in low-iodine well seawater. In collaboration with Israeli counterparts, a highly specific and precise radioimmunoassay procedure for thyroid hormones and cortisol was developed. Taken altogether, the combined Hawaiian and Israeli collaborative research suggests that for teleost species of commercial value, adequate levels of environmental iodine are more determinate in metamorphosis than iodine levels in the live zooplankton food provided to the larvae. Insuring sufficiently high enough iodine in the ambient seawater offers a much more economical solution to improved metamorphosis than enriching the live food with costly liposomes incorporating iodine rich oils.
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Grumet, Rebecca, i Benjamin Raccah. Identification of Potyviral Domains Controlling Systemic Infection, Host Range and Aphid Transmission. United States Department of Agriculture, lipiec 2000. http://dx.doi.org/10.32747/2000.7695842.bard.
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Potyviruses form one of the largest and most economically important groups of plant viruses. Individual potyviruses and their isolates vary in symptom expression, host range, and ability to overcome host resistance genes. Understanding factors influencing these biological characteristics is of agricultural importance for epidemiology and deployment of resistance strategies. Cucurbit crops are subject to severe losses by several potyviruses including the highly aggressive and variable zucchini yellow mosaic virus (ZYMV). In this project we sought to investigate protein domains in ZYMV that influence systemic infection and host range. Particular emphasis was on coat protein (CP), because of known functions in both cell to cell and long distance movement, and helper component-protease (HC-Pro), which has been implicated to play a role in symptom development and long distance movement. These two genes are also essential for aphid mediated transmission, and domains that influence disease development may also influence transmissibility. The objectives of the approved BARD project were to test roles of specific domains in the CP and HC-Pro by making sequence alterations or switches between different isolates and viruses, and testing for infectivity, host range, and aphid transmissibility. These objectives were largely achieved as described below. Finally, we also initiated new research to identify host factors interacting with potyviral proteins and demonstrated interaction between the ZYMV RNA dependent RNA polymerase and host poly-(A)-binding protein (Wang et al., in press). The focus of the CP studies (MSU) was to investigate the role of the highly variable amino terminus (NT) in host range determination and systemic infection. Hybrid ZYMV infectious clones were produced by substituting the CP-NT of ZYMV with either the CP-NT from watermelon mosaic virus (overlapping, but broader host range) or tobacco etch virus (TEV) (non- overlapping host range) (Grumet et al., 2000; Ullah ct al., in prep). Although both hybrid viruses initially established systemic infection, indicating that even the non-cucurbit adapted TEV CP-NT could facilitate long distance transport in cucurbits, after approximately 4-6, the plants inoculated with the TEV-CPNT hybrid exhibited a distinct recovery of reduced symptoms, virus titer, and virus specific protection against secondary infection. These results suggest that the plant recognizes the presence of the TEV CP-NT, which has not been adapted to infection of cucurbits, and initiates defense responses. The CP-NT also appears to play a role in naturally occurring resistance conferred by the zym locus in the cucumber line 'Dina-1'. Patterns of virus accumulation indicated that expression of resistance is developmentally controlled and is due to a block in virus movement. Switches between the core and NT domains of ZYMV-NAA (does not cause veinal chlorosis on 'Dina-1'), and ZYMV-Ct (causes veinal chlorosis), indicated that the resistance response likely involves interaction with the CP-NT (Ullah and Grumet, submitted). At the Volcani Center the main thrust was to identify domains in the HC-Pro that affect symptom expression or aphid transmissibility. From the data reported in the first and second year report and in the attached publications (Peng et al. 1998; Kadouri et al. 1998; Raccah et al. 2000: it was shown that: 1. The mutation from PTK to PAK resulted in milder symptoms of the virus on squash, 2. Two mutations, PAK and ATK, resulted in total loss of helper activity, 3. It was established for the first time that the PTK domain is involved in binding of the HC-Pro to the potyvirus particle, and 4. Some of these experiments required greater amount of HC-Pro, therefore a simpler and more efficient purification method was developed based on Ni2+ resin.